Genetically Engineered Adipose Mesenchymal Stem Cells Using HIV-Based Lentiviral Vectors as Gene Therapy for Autoimmune Diseases

The immunomodulatory and self-renewable features of human adipose mesenchymal stem cells (hAD-MSCs) mark their importance in regenerative medicine. Interleukin 23 (IL- 23) as a proinflammatory cytokine suppresses T regulatory cells (Treg) and promotes the response of T helper 17 (Th17) and T helper 1 (Th1) cells. This pathway starts inflammation and immunosuppression in several autoimmune diseases. The current study for producing recombinant IL- 23 decoy receptor (RIL- 23R) using hAD-MSCs as a good candidate for ex vivo cell-based gene therapy purposes reducing inflammation in autoimmune diseases. hAD-MSCs was isolated from lipoaspirate and then characterized by differentiation. RIL- 23R was designed and cloned into a pCDH-813A- 1 lentiviral vector. The transduction of hAD-MSCs was performed at MOI (multiplicity of infection) = 50 with pCDH- EFI α- RIL- 23R- PGK copGFP. Expressions of RIL- 23R and octamer-binding transcription factor 4 (OCT- 4) were determined by real-time polymerase chain reaction (real time-PCR). Self-renewing properties were assayed with OCT- 4. Bioactivity of the designed RIL- 23R was evaluated by IL- 17 and IL- 10 expression of mouse splenocytes. Cell differentiation confirmed the true isolation of hAD-MSCs from lipoaspirate. Restriction of the enzyme digestion and sequencing verified the successful cloning of RIL- 23R in the CD813A-1 lentiviral vector. The green fluorescent protein (GFP) positive transduction rate was up to 90%, and real-time PCR showed the expression level of RIL-23R. Oct-4 had a similar expression pattern with nontransduced hAD-MSCs and transduced hAD-MSCs/ RIL-23R indicating that lentiviral vector did not affect hAD-MSCs characteristics. Downregulation of IL-17 and upregulation of IL-10 showed the correct activity of the engineered hAD-MSCs. The results showed that the transduced hAD-MSCs/ RIL- 23R, expressing IL-23 decoy receptor, can give a useful approach for a basic research on cell-based gene therapy for autoimmune disorders.

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Genetically Engineered Mesenchymal Stem Cells for Cell and Gene Therapy

Mesenchymal Stem Cell Therapy ◽

10.1007/978-1-62703-200-1_15 ◽

2012 ◽

pp. 321-354

Author(s):

Yunjoon Jung ◽

Jan A. Nolta

Keyword(s):

Stem Cells ◽

Gene Therapy ◽

Mesenchymal Stem Cells ◽

Genetically Engineered ◽

Cell And Gene Therapy

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Mesenchymal Stem Cells Engineered by Nonviral Vectors: A Powerful Tool in Cancer Gene Therapy

Pharmaceutics ◽

10.3390/pharmaceutics13060913 ◽

2021 ◽

Vol 13 (6) ◽

pp. 913

Author(s):

Yuan Ding ◽

Chenyang Wang ◽

Zhongquan Sun ◽

Yingsheng Wu ◽

Wanlu You ◽

...

Keyword(s):

Stem Cells ◽

Gene Therapy ◽

Mesenchymal Stem Cells ◽

Current Knowledge ◽

Cancer Gene Therapy ◽

Genetically Engineered ◽

Bioactive Molecules ◽

Cancer Gene ◽

Nonviral Vectors

Due to their “tumor homing” and “immune privilege” characteristics, the use of mesenchymal stem cells (MSCs) has been proposed as a novel tool against cancer. MSCs are genetically engineered in vitro and then utilized to deliver tumoricidal agents, including prodrugs and bioactive molecules, to tumors. The genetic modification of MSCs can be achieved by various vectors, and in most cases viral vectors are used; however, viruses may be associated with carcinogenesis and immunogenicity, restricting their clinical translational potential. As such, nonviral vectors have emerged as a potential solution to address these limitations and have gradually attracted increasing attention. In this review, we briefly revisit the current knowledge about MSC-based cancer gene therapy. Then, we summarize the advantages and challenges of nonviral vectors for MSC transfection. Finally, we discuss recent advances in the development of new nonviral vectors, which have provided promising strategies to overcome obstacles in the gene modulation of MSCs.

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1417: Mesenchymal Stem Cells Alone or Modified with ENOS Restore Erectile Function in the Aged Rat: Cell-Based Gene Therapy for Erectile Dysfunction

The Journal of Urology ◽

10.1016/s0022-5347(18)38642-7 ◽

2004 ◽

Vol 171 (4S) ◽

pp. 373-373

Author(s):

Trinity J. Bivalacqua ◽

Mustafa F. Usta ◽

Hunter C. Champion ◽

Weiwen Deng ◽

Philip J. Kadowitz ◽

...

Keyword(s):

Stem Cells ◽

Gene Therapy ◽

Mesenchymal Stem Cells ◽

Erectile Dysfunction ◽

Erectile Function ◽

Aged Rat

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Influence of anthocyanins on the adipogenic and chondrogenic differentiation of human adipose mesenchymal stem cells

Intrinsic Activity ◽

10.25006/ia.5.s2-a2.17 ◽

2017 ◽

Vol 5 (Suppl. 2) ◽

pp. A2.17

Author(s):

Liga Saulite

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Chondrogenic Differentiation ◽

Adipose Mesenchymal Stem Cells

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Biological Aspects and Clinical Applications of Mesenchymal Stem Cells: Key Features You Need to be Aware of.

Current Pharmaceutical Biotechnology ◽

10.2174/1389201021666200907121530 ◽

2020 ◽

Vol 21 ◽

Author(s):

Mohammad Saeedi ◽

Muhammad Sadeqi Nezhad ◽

Fatemeh Mehranfar ◽

Mahdieh Golpour ◽

Mohammad Ali Esakandari ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cancer Cells ◽

Immune Modulation ◽

Adult Stem Cells ◽

Inflammatory Diseases ◽

High Capacity ◽

Genetically Engineered ◽

Cartilage Cells ◽

Biological Aspects

: Mesenchymal stem cells (MSCs), a form of adult stem cells, are known to have a self-renewing property and the potential to specialize into a multitude of cells and tissues such as adipocytes, cartilage cells, and fibroblasts. MSCs can migrate and home to the desired target zone where inflammation is present. The unique characteristics of MSCs in repairing, differentiation, regeneration, and its high capacity of immune modulation has attracted tremendous attention for exerting them in clinical purposes, as they contribute to tissue regeneration process and anti-tumor activity. The MSCs-based treatment has demonstrated remarkable applicability towards various diseases such as heart and bone malignancies, and cancer cells. Importantly, genetically engineered MSCs, as a state-of-the-art therapeutic approach, could address some clinical hurdles by systemic secretion of cytokines and other agents with a short half-life and high toxicity. Therefore, understanding the biological aspects and the characteristics of MSCs is an imperative issue of concern. Herein, we provide an overview of the therapeutic application and the biological features of MSCs against different inflammatory diseases and cancer cells. We further shed light on MSCs physiological interaction, such as migration, homing, and tissue repairing mechanisms with different healthy and inflamed tissues.

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Ghrelin peptide improves glial conditioned medium effects on neuronal differentiation of human adipose mesenchymal stem cells

Histochemistry and Cell Biology ◽

10.1007/s00418-021-01980-3 ◽

2021 ◽

Author(s):

Cristina Russo ◽

Giuliana Mannino ◽

Martina Patanè ◽

Nunziatina Laura Parrinello ◽

Rosalia Pellitteri ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Conditioned Medium ◽

Neural Differentiation ◽

Conditioned Media ◽

Glial Fibrillary Acid Protein ◽

Ghrelin Receptor ◽

Neuronal Markers ◽

Adipose Mesenchymal Stem Cells ◽

Marker Expression

AbstractThe influences of ghrelin on neural differentiation of adipose-derived mesenchymal stem cells (ASCs) were investigated in this study. The expression of typical neuronal markers, such as protein gene product 9.5 (PGP9.5) and Microtubule Associated Protein 2 (MAP2), as well as glial Fibrillary Acid Protein (GFAP) as a glial marker was evaluated in ASCs in different conditions. In particular, 2 µM ghrelin was added to control ASCs and to ASCs undergoing neural differentiation. For this purpose, ASCs were cultured in Conditioned Media obtained from Olfactory Ensheathing cells (OEC-CM) or from Schwann cells (SC-CM). Data on marker expression were gathered after 1 and 7 days of culture by fluorescence immunocytochemistry and flow cytometry. Results show that only weak effects were induced by the addition of only ghrelin. Instead, dynamic ghrelin-induced modifications were detected on the increased marker expression elicited by glial conditioned media. In fact, the combination of ghrelin and conditioned media consistently induced a further increase of PGP9.5 and MAP2 expression, especially after 7 days of treatment. The combination of ghrelin with SC-CM produced the most evident effects. Weak or no modifications were found on conditioned medium-induced GFAP increases. Observations on the ghrelin receptor indicate that its expression in control ASCs, virtually unchanged by the addition of only ghrelin, was considerably increased by CM treatment. These increases were enhanced by combining ghrelin and CM treatment, especially at 7 days. Overall, it can be assumed that ghrelin favors a neuronal rather than a glial ASC differentiation.

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